Objective This study aimed to examine the role of long non-coding RNA PCED1B antisense RNA 1(PCED1B-AS1)in the development of hepatocellular carcinoma(HCC).Methods A total of 62 pairs of HCC tissues and adjacent non-t...Objective This study aimed to examine the role of long non-coding RNA PCED1B antisense RNA 1(PCED1B-AS1)in the development of hepatocellular carcinoma(HCC).Methods A total of 62 pairs of HCC tissues and adjacent non-tumor tissues were obtained from 62 HCC patients.The interactions of PCED1B-AS1 and microRNA-34a(miR-34a)were detected by dual luciferase activity assay and RNA pull-down assay.The RNA expression levels of PCED1B-AS1,miR-34a and CD44 were detected by RT-qPCR,and the protein expression level of CD44 was determined by Western blotting.The cell proliferation was detected by cell proliferation assay,and the cell invasion and migration by transwell invasion assay.The HCC tumor growth after PCED1B-AS1 was downregulated was determined by in vivo animal study.Results PCED1B-AS1 was highly expressed in HCC tissues,which was associated with poor survival of HCC patients.Furthermore,PCED1B-AS1 interacted with miR-34a in HCC cells,but they did not regulate the expression of each other.Additionally,PCED1B-AS1 increased the expression level of CD44,which was targeted by miR-34a.The cell proliferation and invasion assay revealed that miR-34a inhibited the proliferation and invasion of HCC in vitro,while CD44 exhibited the opposite effects.Furthermore,PCED1B-AS1 suppressed the role of miR-34a.Moreover,the knockdown of PCED1B-AS1 repressed the HCC tumor growth in nude mice in vivo.Conclusion PCED1B-AS1 may play an oncogenic role by regulating the miR-34a/CD44 axis in HCC.展开更多
AIM:To determine the in vitro protective effect of recombinant prominin-1(Prominin-1)+microRNA-29b(P1M29)on N-methyl-D-aspartate(NMDA)-induced excitotoxicity in retinal ganglion cells(RGCs).METHODS:RGC-5 cells were cu...AIM:To determine the in vitro protective effect of recombinant prominin-1(Prominin-1)+microRNA-29b(P1M29)on N-methyl-D-aspartate(NMDA)-induced excitotoxicity in retinal ganglion cells(RGCs).METHODS:RGC-5 cells were cultured,and NMDAinduced excitotoxicity at the range of 100–800μmol/L was assessed using the MTT assay.NMDA(800μmol/L)was selected as the appropriate concentration for preparing the cell model.To evaluate the protective effect of P1M29 on the cell model,Prominin-1 was added at the concentration of 1–6 ng/mL for 48h,and the cell survival was investigated with/without microRNA-29b.After obtaining the appropriate concentration and time of P1M29 at 48h,real-time polymerase chain reaction(PCR)was utilized to detect the relative mRNA expression of vascular endothelial growth factor(VEGF)and transforming growth factor(TGF)-β2.Western blot detection was applied to measure the phosphorylation levels of protein kinase B(AKT)and extracellular regulated protein kinases(ERK)in RGC-5 cells after treatment with Prominin-1.Apoptosis study of the cell model was conducted by flow cytometry for estimating the anti-apoptotic effect of P1M29.Immunofluorescence analysis was used to analyze the expression levels of VEGF and TGF-β2.RESULTS:MTT cytotoxicity assays demonstrated that P1M29 group had significantly higher cell survival rate than Prominin-1 group(P<0.05).Real-time PCR data indicated that the expression levels of VEGF were significantly increased in both Prominin-1 and P1M29 groups compared NMDA and microRNA-29b group(P<0.05),while TGF-β2 were significantly decreased in both microRNA-29b and P1M29 groups compared NMDA and Prominin-1 group(P<0.05).Western blot results showed that both Prominin-1 and P1M29 groups significantly increased the phosphorylation levels of AKT and ERK compared to NMDA and microRNA-29b groups(P<0.05).Flow cytometry analysis revealed that P1M29 could prevent RGC-5 cell apoptosis in the early stage of apoptosis,while immunofluorescence results showed that P1M29 group had higher expression of VEGF and lower expression of TGF-β2 with a stronger green fluorescence than NMDA group.CONCLUSION:Prominin-1 combined with microRNA-29b can provide a suitable therapeutic option for ameliorating NMDA-induced excitotoxicity in RGC-5 cells.展开更多
目的研究miR-21在弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达,探讨miR-21表达与DL-BCL临床病理特征的关系及其在DLBCL发生发展中的意义。方法采用Real-time RT-PCR方法检测36例DLBCL和10例正常淋巴结中miR-21的表...目的研究miR-21在弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达,探讨miR-21表达与DL-BCL临床病理特征的关系及其在DLBCL发生发展中的意义。方法采用Real-time RT-PCR方法检测36例DLBCL和10例正常淋巴结中miR-21的表达,并采用免疫组织化学SP法检测Ki-67、PTEN在DLBCL中的表达。结果miR-21在DLBCL中高表达,36例DLBCL中有14例表达PTEN(38.9%),21例Ki-67≥50%(58.3%)。DLBCL中miR-21表达水平与PTEN蛋白表达呈负相关,其高表达与DLBCL高Ann Arbor分期、高增殖指数(Ki-67≥50%)、国际预后指数(IPI)呈正相关。结论miR-21过表达可能是DLBCL恶性度高的标志,是促进DLBCL肿瘤细胞增殖的重要因素。PTEN可能是miR-21在DLBCL发挥作用的靶标。展开更多
基金supported by the Medical Science and Technology Research Foundation of Guangdong Province(No.A2020559).
文摘Objective This study aimed to examine the role of long non-coding RNA PCED1B antisense RNA 1(PCED1B-AS1)in the development of hepatocellular carcinoma(HCC).Methods A total of 62 pairs of HCC tissues and adjacent non-tumor tissues were obtained from 62 HCC patients.The interactions of PCED1B-AS1 and microRNA-34a(miR-34a)were detected by dual luciferase activity assay and RNA pull-down assay.The RNA expression levels of PCED1B-AS1,miR-34a and CD44 were detected by RT-qPCR,and the protein expression level of CD44 was determined by Western blotting.The cell proliferation was detected by cell proliferation assay,and the cell invasion and migration by transwell invasion assay.The HCC tumor growth after PCED1B-AS1 was downregulated was determined by in vivo animal study.Results PCED1B-AS1 was highly expressed in HCC tissues,which was associated with poor survival of HCC patients.Furthermore,PCED1B-AS1 interacted with miR-34a in HCC cells,but they did not regulate the expression of each other.Additionally,PCED1B-AS1 increased the expression level of CD44,which was targeted by miR-34a.The cell proliferation and invasion assay revealed that miR-34a inhibited the proliferation and invasion of HCC in vitro,while CD44 exhibited the opposite effects.Furthermore,PCED1B-AS1 suppressed the role of miR-34a.Moreover,the knockdown of PCED1B-AS1 repressed the HCC tumor growth in nude mice in vivo.Conclusion PCED1B-AS1 may play an oncogenic role by regulating the miR-34a/CD44 axis in HCC.
基金Supported by Zhejiang Provincial Natural Science Foundation of China(No.LY20H120003).
文摘AIM:To determine the in vitro protective effect of recombinant prominin-1(Prominin-1)+microRNA-29b(P1M29)on N-methyl-D-aspartate(NMDA)-induced excitotoxicity in retinal ganglion cells(RGCs).METHODS:RGC-5 cells were cultured,and NMDAinduced excitotoxicity at the range of 100–800μmol/L was assessed using the MTT assay.NMDA(800μmol/L)was selected as the appropriate concentration for preparing the cell model.To evaluate the protective effect of P1M29 on the cell model,Prominin-1 was added at the concentration of 1–6 ng/mL for 48h,and the cell survival was investigated with/without microRNA-29b.After obtaining the appropriate concentration and time of P1M29 at 48h,real-time polymerase chain reaction(PCR)was utilized to detect the relative mRNA expression of vascular endothelial growth factor(VEGF)and transforming growth factor(TGF)-β2.Western blot detection was applied to measure the phosphorylation levels of protein kinase B(AKT)and extracellular regulated protein kinases(ERK)in RGC-5 cells after treatment with Prominin-1.Apoptosis study of the cell model was conducted by flow cytometry for estimating the anti-apoptotic effect of P1M29.Immunofluorescence analysis was used to analyze the expression levels of VEGF and TGF-β2.RESULTS:MTT cytotoxicity assays demonstrated that P1M29 group had significantly higher cell survival rate than Prominin-1 group(P<0.05).Real-time PCR data indicated that the expression levels of VEGF were significantly increased in both Prominin-1 and P1M29 groups compared NMDA and microRNA-29b group(P<0.05),while TGF-β2 were significantly decreased in both microRNA-29b and P1M29 groups compared NMDA and Prominin-1 group(P<0.05).Western blot results showed that both Prominin-1 and P1M29 groups significantly increased the phosphorylation levels of AKT and ERK compared to NMDA and microRNA-29b groups(P<0.05).Flow cytometry analysis revealed that P1M29 could prevent RGC-5 cell apoptosis in the early stage of apoptosis,while immunofluorescence results showed that P1M29 group had higher expression of VEGF and lower expression of TGF-β2 with a stronger green fluorescence than NMDA group.CONCLUSION:Prominin-1 combined with microRNA-29b can provide a suitable therapeutic option for ameliorating NMDA-induced excitotoxicity in RGC-5 cells.
